Effects of tunicamycin on retinoic acid induced respecification of positional values in regenerating limbs of the larval axolotl, Ambystoma mexicanum.

Urodele amphibians possess a remarkable ability to regenerate limbs following experimental or accidental amputation. Since only those parts of the limb distal to the plane of amputation usually regenerate, this suggests the existence of level-specific positional values within the cells of the limb. Vitamin A and other retinoids respecify the positional values of regenerating limbs such that structures proximal to the actual plane of amputation are formed in the regenerating limb producing proximodistal duplications. Regenerating limbs of larval axolotls (Ambystoma mexicanum) treated with sufficient retinoic acid to induce proximodistal duplication were also treated via implantation with tunicamycin, a drug which blocks the synthesis of glycoproteins by blocking N-glycosylation of proteins. Tunicamycin was shown to inhibit the proximalizing effects of retinoic acid. This indicates that asparagine-linked glycoproteins may be essential to the process through which retinoic acid induces these effects in the regenerating limb and that glycoproteins may be responsible for specifying positional values in regeneration blastema cells.